Jim Waters: The Development of GPC and the First HPLC Instruments
Aug 1, 2005
By: Leslie S. Ettre
LCGC North America
In the first years, Jim's new company was small. He had only five employees and was operating
in the rental basement of the former police station in Framingham, Massachusetts. His aim was
to develop an instrument for anybody who had the need to measure something. In this period, he
developed a hydrometer for the U.S. Air Force to be used in balloon flights, a flame photometer
for detecting toxic gases, and another flame photometer specially built for the Indian Point
Nuclear Power Plant to check possible salt contamination in the water used in the boilers, which
would corrode the heat exchangers; this photometer could also be sold to the Baltimore Gas &
Electric Co. He also developed a differential refractometer, with a 1-mL cell, which found
limited use in process control in chemical plants.
The Breakthrough: GPC
At that time, LC was still in the laboratory stage, using small glass columns and gravity flow of
the eluent liquid. The only essential difference as compared to the technique of M.S. Tswett 60
years earlier was that by now, the separated individual fractions did not remain on the column
packing but were washed out of the column, the column effluent was collected in small fractions,
and the content of each fraction was determined. However, slowly, the method first described in
the early 1940s by Tiselius in Uppsala, Sweden, also gained in use: the column effluent was
continuously monitored, measuring its refractive index, which changed due to the presence and
amount of the individual sample components (3). Learning about this new trend, Jim Waters was
considering modifying his refractometer for LC use. Then, one day in 1961, he was contacted by
John C. Moore, a young scientist at Dow Chemical Co. in Freeport, Texas who wanted a
refractometer with a very small, 0.1-mL cell, to be used at room temperature, with aqueous flow.
Jim Waters built one for Dow without knowing its actual use. Nine months later, Moore had a
new requirement: he now wanted to have a refractometer that could be used with an organic
solvent (o-dichlorobenzene) instead of water flow, and at 135 °C instead of room temperature.
Again, Jim Waters developed the detector, still without knowing about its planned use. Only
after Dow filed a patent application (4) did he learn that Moore needed the refractometer as a
detector for a new method to establish the molecular weight distribution of high polymers: GPC.
The reason for the two different specifications was that at first, Moore investigated polyglycols,
in aqueous solution and at room temperature, followed by the study of polystyrene samples,
which could only be dissolved in organic solvents like o-dichlorobenzene, and the solutions had
to be kept at elevated temperatures.
After some negotiations, Dow granted exclusive license to
Waters Associates on the technique. The task the company faced
was great: in addition to designing and building an instrument,
they also had to master the synthesis of the rigid cross-linked
polystyrene–divinylbenzene (PS–DVB) gel used as the column
packing. Up until then, Moore only produced this in smaller
batches but now, Jim Waters had to scale up its production to be
Figure 1: The laboratory of
able to supply columns with the instruments. For this, they
Waters Associates in 1963,
converted the female detention area of the former police station
established in the female
in Framingham into a laboratory (Figure 1). After some
detention area of the former
experimental work, he was able to upscale the original
Framingham police station.
Right, in forefront, technician laboratory process to 55 gallon drums and set up regular
production of the material with various pore sizes, sold under the
Nicky Anastis is stirring a
batch of the PS-DVB mixture trade name Styragel.
used to synthesize the Styragel
column packing. The reaction It should be noted that Moore's technique for size-exclusion
vessel was fabricated by Jim chromatography also had its antecedent: gel-filtration
chromatography of Porath and Flodin, in Uppsala, Sweden,
Waters from a standard, 55using soft, compressible, hydrophilic cross-linked dextran gels
gallon metal drum.
with the brand name Sephadex (5). However, these gels only
permitted a slow flow of aqueous solution through relatively short columns and could not be
used for the separation of high-molecular-weight macromolecules. Typically, these columns
have been used in the classical way, collecting small-volume fractions of the column effluent,
but actually, in 1962, Jim Waters personally installed a refractometer detector in Porath's
laboratory to monitor the effluent of gel-filtration columns. In contrast to Sephadex, Moore's
controlled-porosity, rigid PS-DVB copolymer beads could also be used with organic solvents
and permitted the separation of sample components with molecular weights ranging from several
thousand to several million; and while permitting fraction collection, column effluent was
continuously monitored with the differential refractometer detector, producing a true
chromatogram.
Figure 2: The
prototype of the
Model GPC100 gelpermeation
chromatograph
built in 1963.
In 1963, Waters produced five prototypes of the new GPC instrument. These
Top left is the
were large, floor-standing units, larger than a kitchen refrigerator. In these
refractometer
prototypes (Figure 2), the column oven (an air thermostat) was made of
optics module,
plywood, with asbestos insulation; this was then changed in the final version to while the small
a metal cabinet. Three of these prototypes went to Dow Chemical, and one each metal box in
to B.F. Goodrich and Mobil Chemical. The final instrument was introduced to the lower left
the public at the 1964 Pittsburgh Conference under the name GPC-100; its price contains a
was $12,500 (Figure 3). By then, Moore's fundamental paper was finally
device
published (6) and in this way, chemists interested in the new technique could
continuously
learn directly from its inventor. (It is interesting to note that it took well over a measuring the
year to publish this paper: Moore submitted the manuscript to the editorial
volume flow
offices of the Journal of Polymer Science in December 1962, but it was
rate of the
published only in the spring of 1964.)
pump. The
Aside from the small-volume differential refractometer, column oven in
key components of the new instrument were the
the back of the
multiport valves for sample introduction and column
left-hand
switching. The eluent flow was pumped from a reservoir module permits
through a heater to degas the solvent and through a filter the installation
to a mixing chamber needed to dump out pumping
of four 4-ft
pulsations. A Milton Roy pump was used in the
long columns.
instrument; Jim Waters designed his own pump only
All the
electronics are
Figure 3: Illustrations later, for high-pressure liquid chromatographs (see
below).
in the rightfrom the GPC 100
hand module,
Instruction Manual in
with a
1964. The production The acceptance of the new instrument and of the new
potentiometer
technique
was
great,
and
in
1964,
the
company
sold
40
model instrument is
recorder at the
units. The new technique really filled a gap: using
housed in a metal
top.
classical
methods,
it
took
three
to
four
weeks'
hard
work
cabinet, though the
to establish the molecular-weight distribution of a single sample. Now,
arrangement of
with the new instrument, one full analysis could be accomplished in
components is the
about an hour and half.
same as in the 1963
prototypes. The pump,
shown top right, was
made by Milton Roy.
Jims differential
refractometer was a
key innovation. Note
that the fluid system
was termed: "Waters
Liquid
Figure 4: Key participants at
Chromatography
the First Waters GPC Seminar,
Assembly."
held in Cleveland, Ohio, in
January 1965. From the left:
Jim Waters, John C. Moore
(Dow, Freeport, Texas), Prof.
Fred V. Billmeyer (RPI) and
At the end of 1964, Larry E. Maley, Waters' marketing manager Larry E. Maley (VP of Sales,
who was instrumental in demonstrating the new instrument to
Waters). They are holding a
prospective customers, had the brilliant idea to organize the first model representing a 100,000
GPC symposium. This was held in Cleveland, Ohio, in January MW linear polymer.
1965. Two plenary lectures were presented by Fred W. Billmeyer, professor of polymer science
at Rensselaer Polytechnic Institute (RPI; Troy, New York) and by John Moore; these were
followed by about 20 papers by users of the instrument and by lengthy discussions (Figure 4).
The symposium was so successful that Waters repeated it in 1966, now at the Fontainebleau
Hotel in Miami Beach (certainly a more hospitable place than Cleveland in the middle of
winter!). Since then, these symposia became regular yearly events.
In subsequent years, Jim Waters further improved the instrument, introducing a number of
improved models, permitting both analytical and semi-preparative work. Without any question,
making size-exclusion (gel-permeation) chromatography a routine laboratory tool for the
characterization of polymers can be attributed to him.
Liquid Chromatography
In the early 1960s, gas chromatography (GC) was the prevailing chromatographic technique (7)
and, as mentioned earlier, LC was still done essentially in the classical way, with very little
change from Tswett's original description of the technique. However, slowly researchers well
versed in the theory and practice of GC realized that changes were needed to make LC
compatible with regard to both speed and resolution. In my discussion of Csaba Horváth's early
activities (1), I dealt with this trend. Jim Waters was also thinking along this line. In an internal
document from this period, he stated that "We believe LC can become a mass market which will
extend far beyond the research laboratory into production, quality control, and clinical testing"
(8). Waters Associates already sold a limited number of their small-volume refractometers to
chromatographers, to monitor the effluent of classical LC columns. Then, in 1965, Jim became
involved in the development of a liquid chromatograph for Shell Development Co.
Shell Development Co. in California had been involved in the development of new
instrumentation for their own use and usually licensed smaller companies to produce these. For
example, in the early stage of GC development, they licensed a short-lived California company,
Hallaikainen Instruments, to build gas chromatographs based upon their design, and Hallaikainen
also sold a few to customers other than Shell (7). The situation was similar with LC. By the mid1960s, Shell had developed an LC system and in 1965, Jim Waters secured a license to construct
instruments based upon this design. Shell's system was based upon liquid–liquid chromatography
(LLC), and Waters built two prototypes using his refractometer detector: one for Shell and a
second for the University of Alberta, in Canada.
Figure 5: The
Model ALC-100
high-pressure
analytical liquid
chromatograph
introduced in
1968. The box on
the right top is the
compartment for
the solvent bottles;
below it are the
control panels for
the refractometer
(left) and UV
(right detectors
and at the bottom
is the pump
compartment). The
left-hand side of
the instrument is
the thermostatted
column
compartment. The
operator is
injecting a sample
with a syringe into
the injection port.
A separate twopen potentiometric
recorder is used to
record the
chromatograms.
Many of the researchers who started to upgrade LC in the 1960s were not
sure which approach to take: follow the GC example and use columns coated
with a (liquid) stationary phase, or utilize a solid stationary phase (i.e., an
adsorbent); in other words, carry out liquid–liquid or liquid–solid
chromatography (LLC or LSC). Shell's approach was the former. However,
Jim soon realized (just as eventually the other pioneers in this early period of
modern LC development) that this was not the right approach. The system
was not stable and the detector drifted widely. Therefore, Jim completely
redesigned the original Shell system. Meanwhile, reports on the work of
other pioneers started to be known and Karl J. Bombaugh, Waters' director
of R&D, learned at the 1966 Chromatography Symposium held in Rome,
Italy, about the approach selected by Csaba Horváth (1) (I was present and
still remember vividly their discussion). As a conclusion, Jim Waters
switched to the use of solid column packing (an adsorbent), also included a
UV detector, and adopted the system for high-pressure operation. The
resulting instrument, the ALC-100 analytical liquid chromatograph — the
first commercial high-pressure liquid chromatograph — was formally
introduced at the 1968 Pittsburgh Conference (Figure 5). It was a benchtop
system equipped with a Milton Roy pump, syringe injection, and two
detectors: the Waters differential refractometer and a UV detector from the
Laboratory Data Control (LDC) Co.
Almost simultaneously with the ALC-100, Jim
designed another instrument for large-scale
operation, for both GPC and LC: the ChromatoPrep (Figure 6). It was a six-foot tall, floorstanding instrument on casters, equipped with
the same two detectors as the ALC-100. It had
a 40-port valve with an integral, time-based
fraction collector.
As mentioned earlier, Waters Associates first Figure 6: The Chromatoused a pump made by another company.
Prep GPC-LC instrument
However, Jim was aware that the pump was the for preparative-scale work,
key component of a liquid chromatograph;
introduced in 1968. The
therefore, he initiated a major project to
column compartment is
develop the M6000 pump announced in
open, showing four columns
December 1972, and formally introduced at the installed. Below the space in
1973 Pittsburgh Conference (Figure 7). It
the middle of the right side
represented a major innovation, providing
of the module is a fraction
pulseless flow at 6000 psi and flow rates
collector, while the two-pen
between 0.1 and 9.9 mL/min.
potentiometric recorder is
installed in the right top.
A milestone in Jim Waters'
involvement in LC occurred in 1972. At that time, Professor Robert
B. Woodward of Harvard University (who received the 1965
Chemistry Nobel Prize) was involved in the synthesis of Vitamin ±2,
but had difficulties in separating two positional isomers of a key
Figure 7: The Model 6000
solvent-delivery system
for liquid chromatographs
introduced in 1973.
intermediary compound along the synthesis. Woodward left for Europe and entrusted Dr. Helmut
Hamberger, one of his principal postdoctoral assistants, with the task of trying to solve the
problem. In turn, Dr. Hamberger contacted Jim, asking whether LC could be of help. In turn, he
took a Model ALC-100 liquid chromatograph to Harvard and, in cooperation with Dr.
Hamberger, was able to accomplish within a week the separation (8). Upon the return of
Professor Woodward the happy pair could surprise him with 200 mg of the pure compound. A
photograph made with Woodward (Figure 8) was used by Waters Associates to promote their
excellence in LC among university professors involved in organic synthesis work. (Prof. J.F.K.
Huber of the University of Amsterdam, The Netherlands, a pioneer in modern LC, was visiting
Waters Associates at that time and came along with Jim for the visit to Harvard. In 1974, Dr.
Huber became professor at the University of Vienna, in Austria.)
By the early 1970s, Waters Associates was firmly established as
the key player in high-performance liquid chromatography
(HPLC) and thus, they rightly trademarked the tagline "The
Liquid Chromatography People" for the company. This slogan
corresponded to the truth: for years they had a dominant role in Figure 8: At Harvard
the field. By the end of the 1970s, when the worldwide LC
University, in 1972. Left to
market was estimated between $100 and 150 million, Waters had right: Dr. Helmut Hamberger
50% of it [9]. In the 25 years since then, the use of LC has
(Harvard), Prof. J.F.K. Huber
undergone a meteoric rise to become the most widely used
(University of Amsterdam),
laboratory technique. At present, the combined LC and LC–MS James L. Waters, and Prof.
markets is estimated as about $3 billion (10). Naturally, the
R.B. Woodward (Harvard). An
relative share of Waters Associates was slowly reduced (the
ALC-100 liquid
competition "grew up"). However, the company is still the world chromatograph can be seen on
leader in this field (10).
the right; the pump
compartment's door is open.
Jim Waters — An Appraisal
The subject of this review was the activities of Jim Waters in establishing GPC as an important
chromatography technique, and in the early development of instrumentation for HPLC. Still, we
cannot finish our discussion without saying a few words about the last three decades. Jim Waters
served as the president of the company until 1972 when Frank Zenie, up to then the head of
manufacturing, took over as the C.E.O. and Jim became chairman of Waters Associates. In 1973,
the company moved to Milford, Massachusetts, greatly expanding its operation. Finally, in
March 1980, it merged with Millipore Corporation but in 1994 it split, again becoming an
independent company. After the merger with Millipore, Jim Waters left the company and since
then has been focusing on venture capital opportunities.
During his 22-year leadership between 1958 and 1980, Waters Associates grew from five to
1100 employees, with a yearly sales volume of close to $100 million. Today, Waters Corporation
and its subsidiaries have around 4000 employees worldwide, and a yearly sales volume
exceeding $1 billion. Its success is a tribute to its founder, who set the company on the road to
becoming the principal player in LC instrumentation.
Acknowledgements
This review of the activities of James L. Waters and the early development of GPC and LC
instrumentation is based upon a number of previous articles (8,9,11–13), as well as on additional
information obtained from Dr. Patrick D. McDonald, Senior Fellow of Waters Corporation, who
has been associated with the company for over three decades. The figures were obtained from
Waters Corporation through Dr. McDonald. I would like to express my appreciation to Dr.
McDonald for his help and to Waters Corporation for permission to use copyrighted material.
Leslie S. Ettre From 1988 to 2004, "Milestones in Chromatography" editor Leslie S. Ettre was
associated with the Chemical Engineering Department of Yale University (New Haven,
Connecticut), first as an adjunct professor and then as a research fellow. Previously, he had
been with the Perkin-Elmer Corporation for 30 years. He is currently a member of LCGC's
editorial advisory board. Direct correspondence about this column to "Milestones in
Chromatography," LCGC, Woodbridge Corporate Plaza, 485 Route 1 South, Building F, First
Floor, Iselin, NJ 08830, e-mail lcgcedit@lcgcmag.com
.
References
(1) L.S. Ettre, LCGC 23(5), 486–495 (2005).
(2) H.A. Liebhafsky, Anal. Chem. 34(7), 23A–33A (1962).
(3) L.S. Ettre, in High-Performance Liquid Chromatography — Advances and Perspectives, vol.
1, Cs. Horváth, ed. (Academic Press, New York, 1980), pp. 1–74.
(4) J.C. Moore, U.S. Patent 3,326,875 (Filed: Jan. 31, 1963; issued: June 20, 1967).
(5) J. Porath and P. Flodin, Nature (London) 183, 1657–1659 (1959).
(6) J.C. Moore, J. Polymer Sci. A2, 835–843 (1964).
(7) L.S. Ettre, J. Chromatogr. Sci. 15, 90–110 (1977).
(8) B.J. Murphy, "About Waters" tab /Waters/Profile/History in: http://www.waters.com
<http://www.waters.com> (2003).
(9) Forbes Magazine, May 1979, pp. 162–164.
(10) "The Laboratory Life Science and Analytical Instrumentation Industry" (Strategic
Directions International, Inc., Los Angeles, California, June 2004), pp. 62–71.
(11) Gordon Wilkinson, Analytical Instrument Industry Report 7(4), 4–5 (1990).
(12) J. Poudrier and J. Maynihan, in Made to Measure (American Chemical Society,
Washington, DC, March 1999), pp. 11–12.
(13) D. O'Reilly, Makers of Modernity — Members of the Pittcon Hall of Fame (Chemical
Heritage Foundation, Philadelphia, Pennsylvania, 2005), p. 28.
Jim Waters: The Development of GPC and the First HPLC Instruments
Figure 3: Illustrations from the GPC 100 Instruction Manual in 1964. The production model instrument is
housed in a metal cabinet, though the arrangement of components is the same as in the 1963
prototypes. The pump, shown top right, was made by Milton Roy. Jims differential refractometer was a
key innovation. Note that the fluid system was termed: "Waters Liquid Chromatography Assembly."
Figure 5: The Model ALC-100 high-pressure analytical liquid chromatograph introduced in 1968. The box
on the right top is the compartment for the solvent bottles; below it are the control panels for the
refractometer (left) and UV (right detectors and at the bottom is the pump compartment). The left-hand
side of the instrument is the thermostatted column compartment. The operator is injecting a sample
with a syringe into the injection port. A separate two-pen potentiometric recorder is used to record the
chromatograms.